Overall objectives are defining proliferation and differentiation of the lymphoid precursors with the most long term repopulating ability, primitive stem cells (PSC). All lymphoid and myeloid cells are continuously descended from PSC. This project focuses on precursors that repopulate the lymphoid system over many months. Their functions are vital to health.
Specific aims are: (1) To determine whether PSC are a distinct class of cells with maximal long term repopulating abilities by competitive repopulation of untreated B6 cells mixed with B6-HbbdGpi-1A cells individually marked by retroviral insertion in collaboration with I.R. Lemischka, limiting dilution in recipients with differing repopulating abilities, and testing whether enrichment separates precursors that only repopulate during a few weeks. (2) To compare effects of replication and migration on transplanted cells, by recovery after irradiation with differing portions of marrow shielded. (3) To eliminate damage from irradiation and optimize stimuli for PSC proliferation, using double W- anemic, scid mutants for repopulation without irradiation, and recipients with differing degrees of stimuli to differentiate. (4) To define early repopulation after engraftment by multipotential and lymphoid specific precursors, testing whether T and B cells, and also myeloid cells, are descended from the same precursor. (5) To identify PSC markers, enrich PSC, and develop PSC-specific monoclonal antibodies, identifying markers by effects on repopulation, and using enriched PSC to produce PSC specific monoclonals in hamsters and in W-anemic mice. Binding cells with known PSC markers that give at least 1500 fold enrichment will be used as a convenient screen. Existing monoclonals will also be tested. PSC populations will be analyzed using new techniques that directly measure repopulating and differentiation ability in vivo over many months, and estimate numbers of the precursors from which most differentiated cells are descended. These measures will be supplemented by unique retroviral insertions and limiting dilution. The former will define duration and extent of function of individual clones of precursors.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK025687-18
Application #
3227531
Study Section
Immunobiology Study Section (IMB)
Project Start
1979-12-01
Project End
1996-11-30
Budget Start
1992-12-01
Budget End
1993-11-30
Support Year
18
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Jackson Laboratory
Department
Type
DUNS #
042140483
City
Bar Harbor
State
ME
Country
United States
Zip Code
04609
Harrison, D E; Zhong, R K; Jordan, C T et al. (1997) Relative to adult marrow, fetal liver repopulates nearly five times more effectively long-term than short-term. Exp Hematol 25:293-7
Zhong, R K; Astle, C M; Harrison, D E (1996) Distinct developmental patterns of short-term and long-term functioning lymphoid and myeloid precursors defined by competitive limiting dilution analysis in vivo. J Immunol 157:138-45
Zhong, R K; Donnenberg, A D; Edison, L et al. (1996) The appearance of Thy-1- donor T cells in the peripheral circulation 3-6 weeks after bone marrow transplantation suggests an extrathymic origin. Int Immunol 8:171-6
Jordan, C T; Astle, C M; Zawadzki, J et al. (1995) Long-term repopulating abilities of enriched fetal liver stem cells measured by competitive repopulation. Exp Hematol 23:1011-5
Harrison, D E; Zsebo, K M; Astle, C M (1994) Splenic primitive hematopoietic stem cell (PHSC) activity is enhanced by steel factor because of PHSC proliferation. Blood 83:3146-51
Gardner, R V; Lerner, C; Astle, C M et al. (1993) Assessing permanent damage to primitive hematopoietic stem cells after chemotherapy using the competitive repopulation assay. Cancer Chemother Pharmacol 32:450-4
Harrison, D E; Jordan, C T; Zhong, R K et al. (1993) Primitive hemopoietic stem cells: direct assay of most productive populations by competitive repopulation with simple binomial, correlation and covariance calculations. Exp Hematol 21:206-19
Harrison, D E; Zhong, R K (1992) The same exhaustible multilineage precursor produces both myeloid and lymphoid cells as early as 3-4 weeks after marrow transplantation. Proc Natl Acad Sci U S A 89:10134-8
Harrison, D E (1992) Evaluating functional abilities of primitive hematopoietic stem cell populations. Curr Top Microbiol Immunol 177:13-30
Harrison, D E; Lerner, C P (1991) Most primitive hematopoietic stem cells are stimulated to cycle rapidly after treatment with 5-fluorouracil. Blood 78:1237-40

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